Regulatory role of l-proline in fetal pig growth and intestinal epithelial cell proliferation

l-proline (Pro) is a precursor of ornithine, which is converted into polyamines via ornithine decarboxylase (ODC). Polyamines plays a key role in the proliferation of intestinal epithelial cells. The study investigated the effect of Pro on polyamine metabolism and cell proliferation on porcine enterocytes in vivo and in vitro. Twenty-four Huanjiang mini-pigs were randomly assigned into 1 of 3 groups and fed a basal diet that contained 0.77% alanine (Ala, iso-nitrogenous control), 1% Pro or 1% Pro + 0.0167% α-difluoromethylornithine (DFMO) from d 15 to 70 of gestation. The fetal body weight and number of fetuses per litter were determined, and the small and large intestines were obtained on d 70 ± 1.78 of gestation. The in vitro study was performed in intestinal porcine epithelial (IPEC-J2) cells cultured in Dulbecco''s modified Eagle medium-high glucose (DMEM-H) containing 0 μmol/L Pro, 400 μmol/L Pro, or 400 μmol/L Pro + 10 mmol/L DFMO for 4 d. The results showed that maternal dietary supplementation with 1% Pro increased fetal weight; the protein and DNA concentrations of the fetal small intestine; and mRNA levels for potassium voltage-gated channel, shaker-related subfamily, member 1 (Kv1.1) in the fetal small and large intestines (P < 0.05). Supplementing Pro to either gilts or IPEC-J2 cells increased ODC protein abundances and polyamine concentrations in the fetal intestines and IPEC-J2 cells (P < 0.05). In comparison with the Pro group, the combined administration of Pro and DFMO reduced the expression of ODC protein and spermine concentration in the fetal intestine, as well as the concentrations of putrescine, spermidine and spermine in IPEC-J2 cells (P < 0.05). Meanwhile, the percentage of cells in the S-phase and the mRNA levels of proto-oncogenes c-fos and c-myc were increased in response to Pro supplementation, whereas depletion of cellular polyamines with DFMO increased tumor protein p53 (p53) mRNA levels (P < 0.05). Taken together, dietary supplementation with Pro improved fetal pig growth and intestinal epithelial cell proliferation via enhancing polyamine synthesis.     

马疱疹病毒1型主要毒力基因分析及TK基因缺失载体的构建

为了解新疆马疱疹病毒1型（EHV-1）主要毒力基因遗传进化情况并构建TK基因缺失株，本研究以EHV-1 XJ2015株DNA为模板，对其主要毒力基因TK、gI和gE全长进行克隆、测序及生物信息学分析，并扩增TK基因左右重组臂TKL和TKR，构建质粒pUC-TKLR，将扩增后的增强绿色荧光蛋白（EGFP，含有CMV+polyA）插入pUC-TKLR质粒，构建TK基因缺失打靶质粒。TK、gI和gE基因同源性分析结果显示，XJ2015株与国外EHV-1分离株TK、gI和gE基因同源性均较高，分别为99.8%~100.0%、99.6%~100.0%和99.9%~100.0%；与EHV-3分离株同源性均最低，分别为72.9%、59.4%和62.1%；遗传进化分析显示，3个基因均与国外EHV-1同属于一个遗传进化分支，与EHV-9和EHV-4进化关系较近，但与EHV-3进化关系较远，表明XJ2015毒株与国外EHV-1毒株TK、gI、gE基因核苷酸上差异不明显，没有明显的地域性特征，功能基因保守且进化缓慢，同源基因功能相同或相近；经PCR扩增、酶切、测序及转染鉴定，本试验成功构建了用于TK基因缺失的打靶质粒pUC-TKLR-EGFP。通过对EHV-1主要毒力基因的分析及TK基因缺失打靶载体的构建，为新疆地区马鼻肺炎流行病学调查分析、TK基因缺失株的构建提供理论依据。     

Influencing in vitro clonal propagation of Chonemorpha fragrans(moon) Alston by culture media strength,plant growth regulators,carbon source and photo periodic incubation

Chonemorpha fragrans is an endangered medicinal woody climber,regarded among alternative plant sources of camptothecin.Camptothecin is a monoterpene indole anti-cancer alkaloid with annual trade value of over three billion U.S.dollars in the recent,and is used in the production of its analog drugs approved for the chemotherapy of cancer of varied types.Effects of plant growth regulators,culture media strength and photoperi-odic duration on the micropropagation ef ciency of C.fragrans from nodal segment explants were studied on Murashige and Skoog(MS)medium amended with Thidiazuron(TDZ),Benzylaminopurine(BAP)or Kinetin(Kin).Thidiazuron was more ef cient over BAP and Kin when half basal MS medium was used over full or quarter strength.Results of carbon source experiment showed sucrose as the most effective over glucose,fructose,and maltose in the clonal production.Studies on the photope-riodic incubation duration showed 12 h as the best light period and sub or supra-optimal resulted in the production of abnormal and albino micro shoots.Experimental results on the evaluation of physiological,biochemical parameters showed the role of pigment molecules and antioxidant systems in the production of albino micro shoots.     

Identification of an H1N1 subtype of swine influenza virus and serological analysis

To investigate the epizootic of swine influenza virus(SIV), 60 nasal swabs were collected from a clinical cases of pig farm in Tai'an City, Shandong Province of China in April 2017. SIV was isolated by inoculating into 10-day-old Special Pathogen Free embryonated eggs and the whole genome was sequenced. An H1N1 subtype SIV was isolated and designated as A/swine/Shandong/TA04/2017(H1N1). Phylogenetic analysis showed that apart from the polymerase A(PA) fragment belonging to the 2009 pandemic H1N1 branch, seven genome segments belonged to avian-like H1N1 influenza virus lineage. The cleavage site sequence of the hemagglutinin(HA) protein was PSIQSR↓G, which is a typical molecular biological characteristic. Five potential N-glycosylation sites(N14, N26, N277, N484 and N543) were found in the HA gene. To further investigate the epidemiology of SIV in this farm, the 995 serum samples were assessed with EAH1N1 2009 pandemic H1N1 and H3 N2 antigens. The results showed that the total positive rate was 65.43%. The positive rates of single virus infection detected by EAH1N1, 2009 pdmH1N1 and H3 N2 for serum HI(Hemagglutination inhibition) were 48.35, 30.85 and 7.47%, respectively. The results showed that SIV in Shandong Province has been reassorted in some segments and the SIV-positive rate was high on the SIV outbreak farm. These data provide evidence of an epizootic of SIV.     

面向养殖水体分布差异的COD光谱法检测研究

面向养殖水体，传统光谱法对化学需氧量（Chemical Oxygen Demand，COD）检测模型构建的基础：源域（现有样本库）与目标域（检测地水体）间光谱数据独立同分布。但是当源域与目标域分布间存在差异时，由源域得到的低误差模型常在目标域上表现下滑。针对该问题，提出面向UV Vis光谱的域对抗训练网络（DAUVwpNet），将分布不同的源域和目标域数据映射至相同分布的特征空间中，使其在该空间的分布距离尽可能接近，从而在特征空间中对源域训练的目标函数也可以迁移至目标域上，以降低模型在目标域的误差。试验表明：面向同一批测试数据，DAUVwpNet的预测误差为0.78，要低于传统模型的预测误差（0.85）；DAUVwpNet预测值与实测值间相关系数为0.95，要高于传统模型的相关系数（0.89）。表明了该网络能够较好对齐两域特征空间数据分布，降低因分布差异带来的COD检测误差。     

大豆质核互作雄性不育系NJCMS1A与其保持系NJCMS1B的生理生化特性比较分析

以大豆质核互作雄性不育系NJCMS1A及其保持系NJCMS1B的花芽为试验材料,利用试剂盒法和紫外分光光度计测定分析了糖分含量(可溶性糖和淀粉)和总ATPase、蔗糖磷酸合成酶(SPS)、磷酸烯醇式丙酮酸羧激酶(PEPCK)、抗坏血酸氧化酶(AO)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)以及过氧化物酶(POD)等酶活性的变化情况。结果表明:相较于保持系NJCMS1B,总ATPase、PEPCK、AO和CAT活性在不育系NJCMS1A中显著下降;糖分含量、SPS和SOD活性在不育系NJCMS1A中下降水平不显著;POD活性在不育系NJCMS1A中显著上升。根据结果分析推测,与能量代谢或胁迫响应等相关的物质或酶活性在不育系NJCMS1A中的亏损现象可能是NJCMS1A花粉败育的主要原因。     

花生栽野种间杂交异源多倍化早期世代基因表达变化分析

为了进一步认识花生种间杂交异源多倍体进化过程中的基因表达变化规律,采用cDNA-HFO-TAG技术,研究花生种间杂交组合(四倍体栽培种‘仲恺花4号’×二倍体野生种Arachis. diogio)杂种F_1和早期多倍体世代S0~S3的基因表达变化情况。14条HFO-TAG引物共扩增出121条cDNA片段,其中差异片段84个,主要包括三种类型:亲本转录物完全沉默(3个),双亲转录物在后代部分材料中沉默(59个)和新转录物激活(22个),上述变化在F1代即开始发生。筛选其中大小为500~2 000 bp的35个TDFs进行克隆测序,有27个和NCBI数据库中已录入的基因具有较高的相似性,包括抗逆相关基因(10条)、未知功能蛋白基因(8条)、能量与代谢相关基因(7条)和转录因子相关的基因(2条)。这些研究结果进一步表明在花生种间杂交异源多倍化早期发生着快速、剧烈的基因表达变化,从中获得的差异基因片段,有助于了解花生属种间杂交异源多倍化早期分子机制变化,这对有效利用野生花生种质优异基因具有重要意义。cDNA-HFO-TAG技术简单、有效且实用,完全适用于花生属基因表达变化研究,可以作为花生属及其它物种基因表达变化分析技术的有效补充。     

保护地黄瓜新品种唐杂6 号的选育

唐杂6 号是以雌性系S16 为母本，以自交系S26 为父本配制的强雌型黄瓜一代杂种。生长势强，商品瓜短棒状， 瓜长12～14 cm，横径4.0～4.3 cm，非特异性环境下雌花率95% 以上，瓜皮嫩绿有光泽，白刺，刺瘤稀小，平均单瓜质量 131.8 g 左右，高抗霜霉病，抗细菌性角斑病，耐白粉病；春保护地栽培平均产量可达8 000 kg·（667 m²） ^-1，秋冬保护地栽 培产量可达6 500 kg·（667 m²） ^-1；适合河北、北京、天津及东北地区春、秋保护地种植。     

米槠人工林土壤微生物群落组成对凋落物输入的响应

全球气候变化显著影响森林凋落物数量,进而会对土壤微生物群落造成影响。本研究以亚热带米槠人工林为研究对象,探究不同凋落物量输入处理(凋落物去除、凋落物加倍、对照)下,森林土壤微生物群落组成的变化。结果表明:与去除凋落物相比,凋落物加倍后0~10 cm土壤铵态氮(NH4^+-N)、硝态氮(NO3^--N)、全氮(TN)、有效磷(AP)含量分别显著增加了30.30%、49.66%、12.77%和13.90%。与对照相比,凋落物加倍与去除处理土壤微生物生物量碳(MBC)和氮(MBN)含量分别显著增加和下降(P<0.05),但凋落物加倍与去除处理间无显著差异。凋落物加倍处理下土壤丛枝菌根真菌(AMF)、革兰氏阳性菌[G(+)]、革兰氏阴性菌[G(-)]、放线菌(ACT)、真菌(F)丰度和总磷脂脂肪酸(TPLFA)含量分别比去除凋落物处理的土壤高68.35%、63.35%、82.65%、69.02%、40.56%和65.85%,而土壤革兰氏阳性菌与阴性菌比值、真菌与细菌比值则分别降低11.64%和26.67%。冗余度分析表明,铵态氮是影响该人工林土壤微生物群落组成的最主要环境因子。可见凋落物输入量变化改变了土壤养分有效性,进而显著影响了土壤微生物群落组成,这对进一步深入探究全球气候变化对亚热带森林土壤养分循环的影响具有重要意义。     

Development of a core set of KASP markers for assaying genetic diversity in Brassica rapa subsp. chinensis Makino

Single‐nucleotide polymorphisms (SNPs) are rapid, economical and reliable genotyping tools. Non‐heading Chinese cabbage (Brassica rapa L. subsp. chinensis Makino) is now an economically important vegetable crop worldwide. In this study, 1,167 SNPs were evaluated for 7polymorphism among 70 representative non‐heading Chinese cabbage inbred lines using a Kompetitive Allele Specific PCR (KASP) genotyping assay. On the basis of identified polymorphisms and the results of a principal component analysis, we selected 50 core SNPs that were balanced sufficiently to provide adequate information for genetic identification. The core SNPs were used for construction of a neighbour‐joining dendrogram that separated the 70 inbred lines into four main groups and several subgroups corresponding to Caixin, Heiyebaicai, Huangxinwu, Naibaicai, Taitsai, Pak‐choi, and Wutatsai. This categorization was superior to that achieved using a dataset of 479 polymorphic SNPs. To confirm the utility of the core SNP markers in genetic identification, we tested their stability and resolution using 162 commercial hybrid cultivars. The SNPs, which represent a cost‐effective, accurate marker set for germplasm analysis and cultivar identification, are suitable for molecular marker‐assisted breeding in non‐heading Chinese cabbage.